Search Results (73,562)

In the present study, a series of isoxazole derivatives were severally evaluated for their antifungal activity against the yeast Candida albicans and molds such as Aspergillus niger, Aspergillus flavus, and Fusarium oxysporum. The results demonstrate that the isoxazole derivatives exhibit considerable antifungal potential, particularly isoxazole-sulfonate ester 4b (Ar= 4-(Cl)C₆H₄, Ar′= 4-(CH₃)C₆H₄), which was found to be active with significant inhibition zones; the diameters of the C. albicans and F. oxysporum samples were measured at 17.00 ± 0.00 mm and 14.00 ± 0.00 mm, respectively. Furthermore, compounds 4a (Ar= 4-(CH₃)C₆H₄, Ar′= 4-(CH₃)C₆H₄), 4c (Ar: 4-(Cl)C₆H₄, Ar′: 4-(NO₂)C₆H₄) and 4d (Ar: 4-(Cl)C₆H₄, Ar′: 3-(Cl)-2-(OCH₃)C₆H₃) demonstrated MIC and MFC values of 20 µg/mL against C. albicans. In addition, the anti-hemolytic activity of these derivatives was evaluated. Compounds 4a, 4e (Ar: 4-(Cl)C₆H₄, Ar′: 3,4-(OCH₃)₂C₆H₃) and aroylisoxazole 3a (Ar: 4-(CH₃)C₆H₄) demonstrated a high degree of anti-hemolytic activity (>99%) at all concentrations evaluated (10, 15, and 20 mg/mL). Molecular docking and molecular dynamics studies over 200 ns revealed protein–ligand complexes to have high affinity and stability, which agrees with the experimental results. The compounds 4d, 4e, and 3a have shown significant interaction with the target proteins of C. albicans, A. flavus, and F. oxysporum, respectively. The results have revealed that the major interaction sites are hydrogen bonding, hydrophobic interactions, and the presence of a water molecule, especially with key residues like TYR_84, ASP_120, SER_90, and THR_89. The crystal structure of compound 4a was also obtained. Full article

Dravet syndrome (DS) is a severe, catastrophic childhood epilepsy predominantly caused by loss-of-function mutations in the SCN1A gene, which encodes the voltage-gated sodium channel Na_v1.1. In this study, we evaluated the therapeutic potential of 5-(Benzofuran-2-yl)-3-(2-chloro-4-fluorobenzyl)-1,3,4-oxadiazol-2(3H)-one (GM-90663), a novel small molecule designed to address the complex pathophysiology of DS. Using scn1lab knockout (KO) zebrafish larvae—a robust vertebrate model for DS—we demonstrated that GM-90663 significantly alleviates seizure-like behavioral movements and rescues deficit in cognitive-like functions. Whole-cell patch-clamp recordings in hippocampal slices revealed that GM-90663 modulates voltage-gated Na⁺ channel kinetics; specifically, it suppresses slow ramp-induced currents, thereby effectively attenuating neuronal hyperexcitability. Furthermore, neurochemical profiling indicated that GM-90663 treatment leads to a marked increase in endogenous serotonin (5-HT) levels in both wild-type and KO larvae. Molecular docking simulations and subsequent in vitro enzymatic assays confirmed that this elevation in serotonin is mediated through the potent inhibition of monoamine oxidase (MAO) activity. Collectively, our findings suggest that GM-90663 exerts its anti-seizure effects through a synergistic dual mechanism—stabilizing sodium channel conductance and elevating serotonergic activity—positioning it as a promising multi-target candidate for the treatment of DS. Full article

Proteasome inhibitors (PIs) are central to multiple myeloma (MM) therapy; however, resistance remains a major clinical challenge, particularly in relapsed/refractory disease. To identify functional mediators of carfilzomib (CFZ) resistance, we performed complementary gain-of-function CRISPR activation and pharmacological screening approaches. These unbiased strategies converged on the E3 ubiquitin ligase MDM2 as a modulator of PI response. MDM2 transactivation enhanced MM cell survival and accelerated recovery following CFZ exposure, supporting a causal role in proteotoxic stress tolerance. Pharmacologic inhibition of MDM2 with NVP-CGM097 synergized with CFZ across multiple PI-sensitive and PI-resistant MM cell lines, irrespective of TP53 status. Mechanistically, MDM2 inhibition induced p21 upregulation, cell-cycle arrest, and reduced c-MYC expression, accompanied by impaired activation of DNA damage response mediators. Genetic silencing of MDM2 phenocopied these effects and increased CFZ sensitivity. Importantly, the combination retained efficacy in MM–stromal co-culture models and in primary patient samples, including cases harboring del(17p), while sparing normal peripheral blood mononuclear cells. Collectively, these findings identify MDM2 as a functional driver of PI resistance and support combined MDM2 and proteasome inhibition as a rational therapeutic strategy in MM, including TP53-deficient contexts. Full article

Diseases caused by Fusarium spp. vary around the world. It is important to determine the causals agents and indigenous antagonists against these pathogens. Thus, this study aimed to (i) determine the pathogens of root rot and yellow leaf disease (RRYLD), (ii) select Trichoderma spp. strains to control the pathogens, and (iii) evaluate methods for preparing the antagonistic fungi. Diseased soil samples were collected from pomelo orchards in Ben Tre province, Vietnam. The experiment isolated 08 Fusarium spp. strains, with the fastest growth in PDA in FP-C16, FP-B18, FP-B16, and FP-B03 (8.33–17.3 mm) on day 4 of culture. They were identified as Fusarium fujikuroi FP-C16, F. verticillioides FP-B18, F. verticillioides FP-B16, and F. incarnatum FP-B03. On the other hand, 25 Trichoderma spp. strains were isolated from the pomelo rhizosphere. Among them, 13 Trichoderma spp. strains showed rapid growth and strong antagonistic activity against two Fusarium spp. strains under laboratory conditions. The two Trichoderma spp. strains TP-C40 and TP-G50 had antagonistic efficiencies against FP-C16 and FP-B16 at 47.7–63.5%. The two selected Trichoderma spp. strains were identified as Trichoderma asperellum TP-C40 and T. yunnanense TP-G50. The two Trichoderma spp. strains TP-C40 and TP-G50 reduced the number of leaves and roots infected by Fusarium spp. Full article

In this study we compared seven wild-type strains of Yarrowia lipolytica as a chassis for aliphatic polyester biodegradation. To this end, we overexpressed the native Lip2 lipase under the strong UASB16-TEF promoter and compared the capability of the engineered strains towards polycaprolactone (PCL) biodegradation. The parental strains are wild types isolated from various sources and geographical regions. Despite the fact that all belong to the Yarrowia lipolytica species, a high diversity among the strains was observed. This study clearly indicates that, for biotechnological applications of the engineered strains, it is crucial to select a proper host for an efficient process. Full article

Background/Objectives: The current study was designed to identify the underlying genetic causes of congenital stationary night blindness (CSNB) in the indigenous consanguineous families from the Southern Punjab region of Pakistan, a population where the inherited retinal disorders are relatively common. Methods: A detailed questionnaire and medical examination were done to check the presence of CSNB in the affected individuals of the enrolled families. Whole-exome sequencing (WES) was performed to identify the pathogenic variants, followed by segregation analyses to confirm the segregation of the identified variants with the disease phenotype in the available affected individuals of the families. Results: We identified two novel and three known pathogenic variants in SAG, GRK1, TRPM1, SLC24A1, and GPR179, having established roles in CSNB. Two novel variants, NM_001252020.1 (p.Gly1020Arg) and NM_001004334.3 (p.Trp508Ter), were identified, and their segregation was confirmed in two families, PKIURP102 and PKIURP564, respectively. NM_002929.3 (p.Arg19Ter) and NM_001301032.1 (p.Phe538CysfsTer23) were the reported variants identified in PKIURP17 and PKIURP528 families, respectively. NM_000541.5 (p.Glu306Ter) was identified in two independent families, PKIURP552 and PKIURP565. Conclusions: Identification of five pathogenic variants in five different genes shows the genetic heterogeneity of CSNB in Pakistani patients. Our findings also expand the mutational spectrum of CSNB in the Pakistani population and may help in the identification of mutational hotspots and may help in the genetic diagnosis of CSNB in consanguineous populations. Full article

Wastewater treatment plants (WWTPs) are key reservoirs for antibiotic resistance genes (ARGs), particularly when linked to mobile genetic elements (MGEs) and specific microbial hosts. We applied Oxford Nanopore long-read sequencing using complementary contig- and read-based approaches to profile the resistome, mobilome, and host taxonomy in influent and effluent samples from WWTPs in Slovakia and Taiwan. Multidrug resistance was the dominant class in all samples, reaching 40.29–60.06% in Taiwanese and 20.00–35.56% in Slovak WWTPs, followed by MLS and tetracycline resistance. These differences reflect country-specific inputs: Taiwanese WWTPs receiving hospital effluent showed higher multidrug resistance, while Slovak WWTPs, fed by municipal and agricultural wastewater, were dominated by tetracycline resistance and Aliarcobacter cryaerophilus. In Taiwan, Acinetobacter baumannii carried multiple ARGs, including msrE and the regulatory gene ompR, co-localized with MGEs on plasmid- and chromosome-associated contigs. Clinically important Enterococcus faecium (Taiwan) and Staphylococcus pseudintermedius (Slovakia), both WHO-priority pathogens, were identified as hosts for MLS and multidrug resistance genes co-localized with MGEs. These findings suggest that integrating contig- and read-based long-read analyses improves ARG compartmentalization, MGE co-localization, and host assignment in wastewater environments beyond either approach alone. Full article

Background: Prophylactic human papillomavirus (HPV) vaccines are crucial for preventing HPV-related cancers. This study aimed to preclinically evaluate a novel recombinant 9-valent HPV vaccine produced in Escherichia coli (E. coli), which targets HPV types 6, 11, 16, 18, 31, 33, 45, 52, and 58, and is based on virus-like particles (VLPs) of the HPV major capsid protein L1. Methods: The molecular weight and purity of HPV L1 protein bands were assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with Coomassie Brilliant Blue staining. The morphology and size distribution of VLPs were characterized using cryo-electron microscopy and DLS. The immunogenicity and durability of the recombinant 9-valent HPV vaccine were evaluated in BALB/c mice and Wistar rats. Mice received single or triple immunizations (2-week intervals) of two vaccine batches or Gardasil^®9 (MSD, USA) control at 1/20 human dose. Antibody responses were monitored via ELISA and pseudovirus neutralization assays over 24 weeks. Rats were administered single or triple immunizations (2-week intervals) of high- (1/10), medium- (1/20), or low-dose (1/40) vaccine or Gardasil^®9 control (1/20), with neutralizing antibodies tracked for 16 weeks. Results: Cryo-electron microscopy and DLS revealed that VLPs of each type appeared as uniformly distributed, spherical or ellipsoidal hollow intact particles with a diameter of approximately 45–65 nm. This vaccine demonstrated robust immunogenicity and long-lasting efficacy in BALB/c mice and Wistar rats, with effects comparable to those of the commercially available vaccine Gardasil^®9. Conclusions: The 9-valent HPV vaccine induces robust and persistent immune responses in mice and rats, strongly supporting further clinical trials. It is expected to be an alternative to marketed vaccines and ease the global supply shortage of 9-valent HPV vaccines. Full article

Magnesium is an essential divalent cation required for adenosine triphosphate (ATP)-dependent reactions, nucleic acid metabolism, and ribosomal stability. Bacteria depend on specialized transport systems to maintain intracellular Mg²⁺ homeostasis as it cannot freely cross the phospholipid bilayer. During infection, host nutritional immunity restricts metal availability, and magnesium limitation within the phagosome compromises bacterial metabolism and stability. This review summarizes the major bacterial magnesium transport systems and their roles in survival and pathogenicity, with an emphasis on Salmonella and extension to clinically relevant ESKAPE pathogens. We focus on the PhoPQ-regulated MgtA, MgtB, and MgtC system, in which low magnesium, acidic pH, and other host-derived signals activate PhoPQ to induce mgt gene expression. MgtA and MgtB act as high-affinity P-type ATPases, whereas MgtC promotes bacterial survival within the intramacrophage environment by inhibiting bacterial F-type ATP synthase through specific interactions with subunit a. We also discuss CorA as a conserved channel for basal Mg²⁺ uptake and MgtE as a Mg²⁺-selective channel whose gating responds to intracellular Mg²⁺ and ATP. Finally, we consider the conservation and variation in these systems across pathogenic bacteria and their potential as therapeutic targets for antimicrobial development. Full article

Miang, a traditional fermented tea produced from Camellia sinensis var. assamica, is of notable cultural and socio-economic relevance in Northern Thailand. Traditionally, the non-filamentous fungi-based process (NFP) in western Lanna uses only young tea leaves, resulting in substantial amounts of mature leaves being discarded as agricultural waste. This study aimed to utilize the mature tea leaves by adapting the filamentous fungi growth-based process (FFP) of eastern Lanna using selected tannin-tolerant microorganisms, including Aspergillus niger MLF3, Cyberlindera rhodanensis P3, and Lactiplantibacillus pentosus A14-6. Study on fermentation dynamics and bioactive compound formation based on a 2-step fermentation process: 3-day solid-state fermentation with A. niger MLF3, followed by 7-day submerged fermentation by co-culture of C. rhodaninsis P3, and L. pentosus A14-6 in 500 mL sterile distilled water at 30 °C. Increased activities of polysaccharide-degrading enzymes and organic acids were clearly observed during solid-state fermentation, while the significant changes in polyphenol, antioxidant, and reducing sugar content in cell-free supernatant (CFS) were found after submerged fermentation. The obtained CFS shows inhibitory effects of 90 ± 2.5% and 95 ± 1.8% on α-glucosidase and α-amylase, respectively. Analysis of CFS by E-tongue and E-nose clearly indicated the influence of microbial mixture on the taste and aroma of the fermented products. These results demonstrate not only a high-yielding strategy for the effective biotransformation of mature tea leaves into functional drink products but also significant implications for reducing agricultural waste. Full article

Thyme (Thymus callieri Borbás ex Velen) and summer savory (Satureja hortensis L.) are aromatic plants from the Lamiaceae family widely used in traditional medicine and the food industry. This study provides a comparative analysis of the phytochemical profiles and biological potential of the essential oils (EOs) of these two plant species from Bulgaria. The chemical composition was determined using GC-MS analysis. Biological evaluation included determination of antioxidant activity (DPPH assay), antimicrobial activity (MIC assay), ex vivo anti-inflammatory effects (IL-1β expression in rat stomach smooth muscle preparations), and in vitro antihemolytic activity. GC-MS analysis identified 16 compounds in T. callieri EO, dominated by p-cymene (46.42%) and thymol (35.80%). In contrast, 17 compounds were identified in S. hortensis EO, with carvacrol (58.81%) and γ-terpinene (22.46%) as major constituents. Both EOs exhibited concentration-dependent antioxidant activity, with S. hortensis showing higher radical scavenging potential. In antimicrobial tests, both oils demonstrated broad-spectrum efficacy with MIC values ranging from 0.313 to 2.5 mg/mL. Ex vivo experiments revealed that T. callieri EO significantly increased IL-1β expression, suggesting immune activation, while S. hortensis EO showed a lower effect, indicating higher anti-inflammatory potential. Furthermore, S. hortensis EO demonstrated superior erythrocyte membrane stabilization (antihemolytic activity) compared to T. callieri EO and the reference anti-inflammatory drug Aspirin. In conclusion, the findings highlighted the distinct biological potential of both Bulgarian EOs, suggesting their diverse applicability as natural bioactive agents in the pharmaceutical and food industries. Full article

We evaluated the antimicrobial and antioxidant capabilities of a bacteriocin purified from a recently identified marine Lactococcus lactis (L. lactis) NAN6399 strain, a lactic acid bacterium recovered from Mediterranean coastal waters near Alexandria, Egypt, and identified by combined API 50 CHL phenotypic profiling and 16S rRNA gene sequencing. Bacteriocin purification was achieved by sequential ammonium sulfate precipitation and reverse-phase high-performance liquid chromatography (RP-HPLC). The purified bioactive fraction had an approximate molecular weight of 20 kDa by SDS-PAGE and a 106-amino-acid N-terminal sequence that, upon BLAST alignment, returned 98.1% overall identity to the Lactococcin 972 family bacteriocin AAK06118.1 from L. lactis IL1403, with divergence confined exclusively to the terminal two C-terminal residues. This sequence is structurally and functionally distinct from canonical Lcn972 (L. lactis IPLA 972): the two peptides share an identical 25-residue signal peptide but diverge entirely in their mature bioactive domains, which exhibit only 9.1% sequence identity. Canonical Lcn972 operates through Lipid II-mediated septum disruption and inhibits only Lactococcus species; the NAN6399 peptide, correctly designated as a novel member of the Lcn972-like peptide family, demonstrated broad-spectrum antimicrobial efficacy against multiple indicator organisms (Staphylococcus aureus, Salmonella typhimurium, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Enterococcus faecalis), producing inhibition zones of up to 30 mm and minimum inhibitory concentration (MIC) values as low as 1.25 μg/mL against S. aureus. Antioxidant capacity was assessed using the DPPH radical scavenging assay, with the purified preparation achieving 73.14 ± 0.34% inhibition. Collectively, these data establish L. lactis NAN6399 as the producer of a bifunctional Lcn972-family bacteriocin with both antimicrobial and antioxidant potential, provide the first experimental characterization of the antimicrobial activity of this Lcn972-family branch, and highlight marine LAB as a productive reservoir for novel bioactive peptide discovery. Full article

Climate change is increasingly recognized as a source of psychological distress, yet the prevalence, predictors, and behavioral implications of climate anxiety remain unevenly understood. This study examines climate anxiety, its key drivers, and associated behavioral responses in a multi-country sample of adults. A cross-sectional online survey was conducted across 21 countries using the Climate Change Anxiety Scale (CCAS), alongside measures of awareness, coping strategies, social support, and food-related behaviors, including food waste reduction, increased plant-based food consumption, and home or community gardening. Analyses included descriptive statistics, exploratory factor analysis (EFA), and multivariable regression. Given the uneven country-level representation, results are reported as pooled patterns with a few exploratory cross-country comparisons. Climate anxiety was widely reported, with over 60% of participants indicating that climate challenges were emotionally overwhelming. Regression analyses showed that climate awareness and frequency of climate-related thinking were positively associated with higher anxiety, although the effect sizes were small and explanatory power was limited (R² = 0.055). EFA identified two related dimensions: cognitive concern about future impacts and affective distress. Climate anxiety across countries showed modest variation (2.44–3.23) and no statistically significant differences, despite variation in awareness. A gap between concern and climate action was evident: only 39.1% reported environmentally motivated dietary changes. Cost, limited availability, and lack of information were the main barriers to climate action, and only 24.4% reported frequent social support. These findings indicate that climate anxiety is shaped by both psychological and structural factors, and that reducing it requires not only increasing awareness but also enabling conditions that support meaningful climate action. Full article

Pseudopodospermum szowitzii (DC.) Kuth. a member of the Asteraceae family, grows naturally in the Irano-Turanian phytogeographical area, including Anatolia. In Anatolia, P. szowitsii, known as “goftigoda,” has edible young leaves and roots and is used in folk medicine for antidiabetic and analgesic properties. Nine compounds, including chlorogenic acid and derivatives from the ethyl acetate layer as well as 2,4,6-trimethoxyphenyl-1-O-β-glucopyranoside, 2,4,6-trimethoxyphenyl-1-O-β-apiofuranosyl-(1→6)-β-glucopyranoside, esculetin 6-O-β-glucopyranoside, and esculetin 6-O-β-apiofuranosyl-(1→6)-β-glucopyranoside from the water part of the methanolic extract, were isolated as known compounds. Notably, all esculetin derivatives have been isolated from the Pseudopodospermum for the first time, and among them, three compounds, esculetin 6-O-β-xylosyl-(1→6)-β-glucopyranoside, esculetin 6-O-β-glucopyranosyl-(1→3)-β-glucopyranoside, and esculetin 6-O-β-glucopyranosyl-(1→6)-β-glucopyranoside, were isolated as new esculetin heterosides that have not yet been isolated from any natural sources. The antioxidant activities of the total extract, phases, fractions, and compounds of P. szowitsii were also tested by evaluating their radical-scavenging capacities against DPPH and ABTS radicals. The ethyl acetate phase and the isolated compounds displayed significant antioxidant activity. The most active compound was caffeic acid, with IC₅₀ values of 2.7 µg/mL and 3.4 µg/mL against DPPH and ABTS radicals, respectively, followed by dicaffeoylquinic acid derivatives and their methyl esters. On the other hand, none of the coumarin derivatives exhibited significant radical-scavenging activity. Full article

Although RNA cytosine-5 methylation (m⁵C) is an important post-transcriptional regulatory mechanism, its contribution to plant antiviral immunity remains unclear. In this study, we identified Thiamine thiazole synthase 2 (TaTHI2) as a host mRNA target of the wheat m⁵C methyltransferase TaNSUN2 during infection by Chinese wheat mosaic virus (CWMV), a soil-borne virus that poses a major threat to wheat production. TaNSUN2 contributes to the m⁵C modification of TaTHI2 transcripts, enhancing mRNA stability and sustaining TaTHI2 accumulation. The disruption of a key m⁵C site markedly reduced methylation, weakened TaNSUN2–RNA binding, and accelerated transcript decay, leading to the compromised production of reactive oxygen species (ROS) and increased viral infection. Mechanistically, the TaNSUN2-dependent m⁵C modification stabilized TaTHI2 mRNA, thereby promoting ROS-mediated antiviral defense. Collectively, our results establish the m⁵C modification of TaTHI2 mRNA as a critical post-transcriptional control point in CWMV resistance and highlight TaNSUN2-dependent RNA methylation as an integral component of host antiviral immunity. Full article